Studies On The Establishment Of Efficient Shoot Regeneration System And The Agrobacterium-mediated Transformation Of Vitreoscilla Hemoglobin Gene(vgb) In Lavandula Angustifolia Mill.

Lavandula angustifolia Mill.is one of the most important aromatic plant.SeveralLavandula species are not cultivated worldwide,because Lavander can not grow better inwaterlogging and shadow,in order to use it better for cosmetics and garden,it is a short-cutway to Lavandula species were improved by the transgene.The objective of this study is totransfer the vgb gene to Lavender and to obtain new types of Lavender with characteristic ofproductive and resisting waterlogging through the study of regeneration system andtransformation system mediated by Agrobacterium tumefaciens of Lavender,and established afoundation of wide application of improving Lavender cultivars applying transgenic method.The results of the research are given below:1 .Study of effcient regeneration systemThrough the study of two Lavender cultivars on regeneration system,the author obtainedthe efficient regeneration system of English Lavender and Dentata hybrid Lavender matureleaves. After the leaves were sterilized in 2 minutes by 0.1% Hgcl,then back side of leavestouched medium were cultured on 1/3MS medium supplemented with 1.0mg.L-1 6-BA,1/2MS medium supplemented with 1.0mg.L-1 6-BA.The regeneration frequency of EnglishLavender is 80.3%, the regeneration frequency of Dentata hybrid Lavender is 70.33%in thismedium. When the shoots grow to 2cm high, they were cut from leaves, then transferred on1/2MS +1.0 mg.L-1IAA , the regeneration shoots produce roots early,and the roots are tidyand thick.Because of different genotype , Lavender regeneration potency is different.Theshoot regeneration frequency of English Lavender was higner than Dentata hybrid Lavender.2 .The construction of vgb vectorsThe transformants were obtained after PGBI4ASVHB was introduced intoAgrobacterium tumefaciens EHA105 strain through the method of freeze-melting.3 .Construction of the transgenic system of English Lavender mature leavesAccording to studies on the sterilization concentration of Cef, the ascertainment of Kanscreening pressure,and the effect factors of transform frequence,the transgenic system withAgrobacterium tumefaciens was constructed: the leaf segments without pre-cultured should beinfected 10min in Agrobacterium tumefaciens with OD600=0.4,co-cultivated 1 days,andscreened with 100mg/L Cef for sterilizing Agrobaclerium tomefaciens, 30mg/L Kan forselecting resistant leaves and 10mg/L Kan for resisted shoots.4 .PCR identincation of transgenic plantsThe DNA of transgenic plants were amplified by PCR,the result of electrophoresis wasthat some plants get the same line as the positive control vgb (450 bp).It showed on molecularlevel that the vgb gene had been integrated into the geonome of Lavender.Transgenic plants ofvgb had been obtained,the transformation rate is 1.33%.